Immunofluorescent detection of ADD (acidic domain deleted mutant of ALT-2) protein

Copyright information:Taken from "Heterologous expression of the filarial nematode gene products reveals their potential to inhibit immune function"BMC Biology 2005;3():8-8.Published online 23 Mar 2005PMCID:PMC555940.Copyright © 2005 Gomez-Escobar et al; licensee BioMed Central Ltd. Paraformaldehyde-fixed wild-type promastigotes and cloned transfected parasites were incubated with mouse antiserum to anti-ALT-2, and detected with an anti-mouse-FITC secondary antibody. Cloned transfected parasites were also incubated with normal mouse serum. Bone marrow-derived macrophages were infected with stationary phase amastigotes of wild-type or transfected and parasites at a ratio of 10 parasites per cell. After 7 days, cells were stained with Giemsa. The percentage of macrophages infected with parasites was determined by counting 3 samples (from 3 different wells) of 100 cells. The χtest showed significant differences in the number of infected macrophages between wild-type and transfectants (p < 0.001), but not between wild-type and -transfected parasites.